Isabelle Fayt

Gene expression profile for ectopic versus eutopic endometrium provides new insights into endometriosis oncogenic potential.

Endometriosis is a common gynecological disorder characterized by pain and infertility, where the lesions disseminate everywhere in the body with a preference for the pelvis. In that, it could be regarded as a benign metastatic disease, because its issue is not fatal. However, the molecular bases of this intriguing clinical condition are not well known. The objective of this study is to characterize the transcriptome differences between eutopic vs. ectopic endometrium with a special interest in pathways involved in cancerogenesis. We performed two hybridizations in technical replicate on highly specific long oligonucleotides microarrays (NimbleGen), with cDNA prepared from six-patients pools, where the same patient provided both eutopic and ectopic endometrium (endometriomas). To confirm the expression microarrays data, quantitative RT-PCR validation was performed on 12 individuals for 20 genes. Over 8000 transcripts were significantly modified (more than twice) in the lesions corresponding to 5600 down- or up-regulated genes. These were clustered through DAVID Bioinformatics Resources into 55 functional groups. The data are presented in a detailed and visual way on 24 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways implemented with induction ratios for each differentially expressed gene. An outstanding control of the cell cycle and a very specific modulation of the HOX genes were observed and provide some new evidence on why endometriosis only very rarely degenerates into cancer. The study constitutes a noteworthy update of gene profiling in endometriosis, by delivering the most complete and reliable list of dysregulated genes to date.

Herpesvirus‐like DNA sequences and Kaposi's sarcoma: Relationship with epidemiology, clinical spectrum, and histologic features

The evidence of an infectious agent other than human immunodeficiency virus (HIV) acting as a possible etiologic cause of Kaposis sarcoma (KS) has received considerable attention in the last years. Recently, DNA sequences from a new herpesvirus (HHV‐8) have been observed in several cases of KS. The discovery suggests that this virus may play a role in the pathogenesis of KS. To evaluate these results, we determined the frequency of HHV‐8 DNA sequences in 78 specimens of KS according to different epidemiologic origins (sporadic KS: 6, immunosuppressive drug‐associated: 11, and AIDS‐associated: 61), clinical forms (cutaneous: 69, mucocutaneous: 4 and visceral: 5) and histologic variants (early‐patch: 40, late‐plaque or nodular: 38).

Proliferating activity in columnar cell lesions of the breast

With the introduction of mammographic screening, columnar cell lesions (CCLs) are observed more and more frequently because they are often associated with microcalcifications. Until now, the proliferative activity of these lesions has not been previously evaluated. Ki67 index was performed by immunohistochemistry in CCLs without atypia [columnar cell change (CCC) n = 20 and columnar cell hyperplasia without atypia (CCH without atypia) n = 20], flat epithelial atypia (FEA DIN1A n = 20), low-grade intraductal carcinoma (DIN1C n = 20), high-grade intraductal carcinoma (DIN 2–3 n = 20). Adjacent terminal duct-lobular unit (TDLU) of normal breast tissue served as control. Ki-67 index is extremely low and close in CCLs without atypia (CCC mean 0.1% and CCH mean 0.76%) and paradoxically is lower than in normal TDLU (mean 2.4%) (p < 0.001). In the FEA, in comparison with normal TDLU and CCLs without atypia, the Ki67 is higher (mean 8.2%) (p < 0.001) but extremely close to those of DIN1C (mean 8.9%) (p = 0.6 NS). Lastly, the Ki67 index is higher in DIN 2–3 (mean 25.4%) than in CCLs without atypia and FEA (p < 0.001). CCLs are disparate lesions having in common cells with columnar configuration but different proliferative characteristics. These data represent findings of biological interest which could help us to better understand these controversial lesions.

Sphingosine pathway deregulation in endometriotic tissues

OBJECTIVE To investigate key genes expression of the sphingosine-1-phosphate pathway in endometriotic tissues. DESIGN A case-control laboratory study. SETTING Tertiary care university hospital. PATIENT(S) A total of 31 women, with (n = 16) and without (n = 15) endometriosis took part in the study. INTERVENTION(S) After surgical excision with pathological analysis, endometrial specimens were obtained from women affected or not by endometriosis. MAIN OUTCOME MEASURE(S) SPHK1-2, SGPP1-2, SGPL1, SPHKAP, and S1PR1-5 messenger RNA expression by quantitative real-time polymerase chain reaction (PCR) in the endometrium of 15 disease-free women, 16 eutopic and 16 ectopic endometrium of endometriosis-affected women. The S1PR1 and S1PR2 expression were further investigated by immunohistochemistry. RESULT(S) The SGPP2 expression was decreased in eutopic and ectopic endometrium of endometriosis-affected women (1.7- and 16.7-fold, respectively). The SGPP1, weakly expressed in healthy endometrium, is up-regulated in endometriosis-affected women (11.9- and 64.7-fold, respectively), but its expression remains low. The SGPL1 expression was decreased in ectopic endometrium (3.3-fold) and SPHKAP expression was increased in ectopic endometrium (112.6-fold) compared with endometrium of disease-free women. In endometriosis-affected women, S1PR3 expression was decreased in eutopic and ectopic endometrium (2.1- and 6.3-fold, respectively); S1PR2 and S1PR1 expression was increased in eutopic (2.5-fold) and ectopic endometrium (2.6-fold). These increases were confirmed at the protein levels by immunohistochemistry. CONCLUSION(S) Expression of the enzymes implicated in the regulation of the sphingosine-1-phosphate level balance and of its receptors is overall heavily deregulated in endometriotic lesions in favor of a decreased sphingosine-1-phosphate catabolism. Our results plead for a role of the sphingosine pathway in establishing and survival of endometriotic lesions.

Estrogen and progesterone receptors in smooth muscle component of deep infiltrating endometriosis

OBJECTIVE To analyze the expression of estrogen (ER) and progesterone (PR) receptors in the smooth muscle component (SMC) of deep infiltrating endometriosis (DIE). DESIGN A prospective clinical and pathologic study of 60 cases of DIE. SETTING University Hospital Department of Gynacology. PATIENT(S) Sixty patients with symptomatic DIE (uterosacral endometriosis n = 14; bladder endometriosis n = 10; colonic endometriosis n = 16; rectovaginal endometriosis n = 20). INTERVENTION(S) Laparoscopic surgery. MAIN OUTCOME MEASURE(S) The expression of ER and PR was studied by immunohistochemistry in the SMC directly around endometriotic foci and at distance (at least >1.5 cm) from them in correlation with proliferative and secretory phases of cycle. RESULTS The ER and PR were present in the SMC of DEI in each location excepting colonic endometriosis where ER were absent. Independently of cycles phases the PR were more abundant than ER. With the exception of rectovaginal endometriosis, where the ER and PR were more abundant in the proliferative than in the secretory phase, in other locations the ER and PR did not differ significantly with cycles phases. Last, if ER and PR were more abundant in SMC around endometriotic foci than at a distance from them. However, the difference was not significant. CONCLUSIONS Our data substantially confirm for the first time that in various forms of DIE, ER and PR are present not only in glands and stroma but also in the smooth muscle major histologic component of this disease.

Hormonal Therapy Deregulates Prostaglandin-Endoperoxidase Synthase 2 (PTGS2) Expression in Endometriotic Tissues

CONTEXT Endometriosis is a common gynecologic condition characterized by an important inflammatory process mediated by the prostaglandin pathway. Oral contraceptives are the treatment of choice for symptomatic endometriotic women. However the effects of oral contraceptives use and prostaglandin pathway in endometriotic women are actually still unknown. OBJECTIVE To investigate the expression of prostaglandin pathway key genes in endometriotic tissue, affected or not by hormonal therapy, as compared with healthy endometrial tissue. DESIGN This was a comparative laboratory study. SETTING This study was conducted in a tertiary-care university hospital. PATIENTS Seventy-six women, with (n = 46) and without (n = 30) histologically proven endometriosis. MAIN OUTCOME MEASURES Prostaglandin-endoperoxidase synthase (PTGS)1, PTGS2, prostaglandin E receptor (PTGER)1, PTGER2, PTGER3, and PTGER4 mRNA levels in endometrium of disease-free women and in eutopic and ectopic endometrium of endometriosis-affected women. PTGS2 expression was further investigated by immunohistochemistry, using specific monoclonal antibodies. PTGS2 expression was analyzed at mRNA and protein levels and correlated with taking hormonal treatment. RESULTS PTGS2 expression was significantly increased in eutopic and ectopic endometrium as compared with healthy tissue (induction of 9.6- and 6.3-fold, respectively; P = .001). PTGS2 immunoreactivity increased gradually from normal endometrium to eutopic and ectopic endometrium (h-score of 96.7 ± 55.0, 128.3 ± 66.1, and 226.7 ± 62.6, respectively, P < .001). PTGER2, PTGER3, and PTGER4 expression increased significantly and gradually from normal to eutopic and ectopic endometrium, whereas PTGER1 remained unchanged. Patients under hormonal treatment had a higher PTGS2 expression at transcriptional and protein levels as compared with those without treatment (P = .002 and P = .025, respectively). CONCLUSIONS Prostaglandin pathway is strongly deregulated in eutopic and ectopic endometrium of women suffering from endometriosis for the benefit of an increased PTGS2 expression. We show for the first time that hormonal treatment appears to enhance even more PTGS2 expression. These results contribute to explain why medical treatment could fail to control endometriosis progression.

Metastasis from apocrine carcinoma of the breast to an endometrial polyp

It is not unusual for extra-genital neoplasms to metastasise to the female genital. However, the metastases are localised mainly in the ovary and vagina and rarely in the uterus. Metastasis of extra-genital tract neoplasms to an endometrial polyp has been described only twice; both cases concerned breast carcinomas, one of lobular type [1] and one of ductal type [7]. We would like to report the first observation of metastasis from an apocrine carcinoma of the breast to an endometrial tamoxifeninduced polyp. The patient was a 70-year-old woman who presented with a 1-month history of metrorrhagia. The patient had, 4 years previously, undergone total left breast mastectomy with ipsilateral axillary lymphadenectomy for an infiltrating apocrine breast carcinoma. The tumour was 20×10×10 mm in size with evidence of metastatic involvement of one axillary lymph node and was, therefore, reported as pT1cN1 in accordance with the 1997 International Union Against Cancer (UICC) staging of breast tumours. Complementary studies, including chest X-ray, hepatic ultrasonography and bone isotopic scan were all negative. Post-surgery treatment consisted of radiotherapy and tamoxifen therapy, although the tumour showed only focal and weak positivity for oestrogen receptors. Follow-up was uneventful for 4 years. However, 4 years later, the patient complained of recurrent metrorrhagia. She underwent hysteroscopy with curettage and abdominal ultrasound. The latter revealed a polypoid mass 1.5 cm in length appended to the uterine dome. Total abdominal hysterectomy with bilateral salpingooophorectomy was performed. Gross examination of the uterus confirmed the presence of a polypoid mass 1.5×1.0×0.5 cm in size attached to the uterine dome. Cervix, myometrium and annexes were grossly unremarkable. The microscopical features of the polyp were typical of those induced by tamoxifen therapy. They showed sparse, irregularly scattered, focally dilated glands, lined by either atrophic or proliferative epithelium. The stroma was diffusely fibrotic with thick-walled, dilated blood vessels. Clusters of large eosinophilic cells similar to those observed in the primary breast carcinoma with abundant granular cytoplasm, globoid nuclei and prominent nucleoli, were visible within the polyp stroma (Fig. 1). In addition, numerous vascular emboli from the same eosinophilic cells were also visible. Immunohistochemically, the tumour cells showed strong positivity for AE1 and AE3 cytokeratins, epithelial membrane antigen (EMA) and GCDFP-15 (Fig. 2). In contrast, oestrogen and progesterone receptors were negative. No other microscopic metastases were found in the adjacent endometrium, myometrium, ovaries, tubes or cervix.

Proliferating cell nuclear antigen distribution in verrucous carcinoma of the skin.

Verrucous carcinoma (VC) of the skin is a rare variety of well‐differentiated squamous cell carcinoma (SCC) characterized by aggressive local growth and a low metastatic potential. These tumours are known to have histological and virological features similar to classic warts or condylomata. The aim of the present study was to map the proliferative compartment in VC (n=7) in comparison with warts (n=10) and typical well‐differtntiated SCC (n=10). The proliferating cells were detected by immunostaining of proliferating cell nuclear antigen (PCNA) in formalin‐fixed, paraffin‐embedded tissue sections, using the commercially available anti‐PCNA monoclonal antibody PC10. Normal epidermis served as a positive control and reference. In VC and warts, the PCNA‐positive cells were principally located at the periphery of lesions, in the basal layer of the tumour islands. In some warts, however, stronger PCNA expressed was noted in the superficial layers, of the lesions corresponding to virus‐infected keratinocytes (koilocytotic cells). In contrast, in SCC, PCNA‐positive cells were randomly scattered throughout the tumours.

Galectin-3 is overexpressed in various forms of endometriosis.

Endometriosis is an enigmatic disease of unknown etiology and pathogenesis, which is defined as the presence of endometrial glands and stroma outside the uterus. The most widely accepted theory to explain endometriosis is probably the transplantation of an endometrial fragment during menstruation to ectopic sites, but the development of endometriosis is extremely complex and includes the adherence to the peritoneal surface and secondary invasion of the underlying tissues. In this study, we have investigated the potential role of galectin-3 (gal-3), a member of a group of carbohydrate-binding proteins, which plays a major role in cell adhesion, migration, angiogenesis, and invasion. The expression of gal-3 has been carried out by immunohistochemistry, according to the different phases of cycle in 50 cases of endometriosis (peritoneal endometriosis: n=10; ovarian endometriosis: n=10; deeply infiltrating endometriosis: n=30) and in 34 cases of eutopic endometrium (10 without endometriosis and 24 with endometriosis). In the proliferative and secretory phases of the cycle, the nuclear and membranous gal-3 expression was higher, first in each variant of the endometriosis than in the eutopic endometrium (P<0.05), and second in the eutopic endometrium of women with endometriosis than in eutopic endometrium of women without endometriosis. Our data suggest that gal-3 may have a potential role in the development of endometriosis.

Overexpression of 27-kDa heat shock protein relates to poor histological differentiation in human oesophageal squamous cell carcinoma.

Various stress conditions such as heat, chemical and mechanical stresses are known to play a major role in oesophageal squamous cell carcinoma development. Our goal was to evaluate whether changes in stress‐induced 27‐kDa heat shock protein (HSP27) expression could be demonstrated during oesophageal carcinogenesis.